Halogenated composition, method for preparing same and uses thereof

ABSTRACT

The invention concerns pharmaceutical composition including (i) at least a halogenated compound and (ii) at least N-halogenated derivative of at least a compound selected from zwitterionic compounds and/or amino acids. The halogenated compound is advantageously an alkaline metal hypochlorite, and preferably sodium hypochlorite, and N-halogenated derivative is preferably an N-halogenated taurine derivative and particularly a taurine N-haloamine derivative and even more preferably taurine N-chloramine. The invention also concerns the preparation of the compositions and their uses as very large spectrum antiseptic, anti-inflammatory agent and as immunity modulator, without stimulating myeloperoxidase activity.

RELATED APPLICATION

[0001] This is a continuation of International Application No.PCT/FR02/00151, with an international filing date of Jan. 16, 2002,which is based on French Patent Application No. 01/00862, filed Jan. 23,2001.

FIELD OF THE INVENTION

[0002] This invention relates a new composition containing halogenatedcompounds for (1) the treatment of viral, bacterial, parasitical, fungalinfections, or infections generated from non-conventional transmissibleagents; (2) the treatment of chronic, progressive or acute inflammation;(3) immuno-modulator treatments, and/or tissue healing stimulatortreatments; and (4) pre- and/or per- and/or post-surgical irrigations.The invention composition is particularly helpful as a local useantiseptic.

BACKGROUND

[0003] 1. The hypochlorite of alkaline metal.

[0004] Hypochlorite of alkaline metal and, particularly, the sodiumhypochlorite (NaOCl), has been used since the 19^(th) century for itsantiseptic properties. Alkaline metal hypochlorite is an alkaline metalsalt of hypochlorous acid. The available chloride level of sodiumhypochlorite solutions is equal to the addition of HOCl (hypochlorousacid) and OCl⁻ (hypochlorous anion) concentrations (Bloomfield & Miles,1979). The hypochlorite active form, i.e., the hypochlorous acid, is ahighly strong oxidant that plays a role in the mammalian defense system.HOCl is synthesized in polymorphonuclear neutrophils and monocytes(Wright et al., 1986) during the respiratory burst by themyeloperoxidase-H₂O₂-halide system. Hypochlorous acid is unstable andreacts readily with primary and secondary amines to generate variousN-chloramines (Zgliczynski et al., 1971).

[0005] In polymorphonuclear cytosol and, especially in neutrophilcytosol, an amino acid (i.e. taurine) is particularly abundant and has avery high reactivity with hypochlorous acid to yield the taurineN-chloramine (TauCl). This chloramine is less toxic and reactive thanhypochlorous acid. In addition, TauCl is the most stable of thechloramines (Zgliczynski et al., 1971; Marquez & Dunford, 1994).Moreover, taurine seems to have a high protective role in both intra-and extra-cellular environments, via its high scavenger activity withhypochlorous acid (Cantin, 1994; J. Marcinkiewicz et al., 1998).However, long-lived taurine N-chloramines can move and react (i.e.,oxidize and/or chlorinate) at distance from their formation to generatetissue damage (Zgliczynski et al., 1971).

[0006] At the physiological pH (7.4), taurine and hypochlorous acidreact spontaneously and with a 1/1-molecule stoechimetry to yield ataurine N-monochloramine. At acidic pH, this reaction generates bothtaurine N-monochloramines and taurine (N,N)-dichloramines. Taurine and,particularly, nitrites (NO₂ ⁻), compete with other antioxidants toscavenge hypochlorous acid in the extracellular medium. Theirconcentrations are roughly equal. Thus, the main hypo-chlorous acidscavengers are nitrites, which react together to yield a lesser toxicderivative than TauCl. In the polymorphonuclear neutrophil cytosol, dueto its high concentration (≈20 mM), taurine is the main scavenger ofhypochlorous acid (J. Marcinkiewicz, 2000).

[0007] 2. Sodium hypochlorite, hypochlorous acid, N-chloramineproperties.

[0008] a. Dissolving tissue abilities.

[0009] In aqueous solution, the sodium hypochlorite (NaOCl) is wellknown to be caustic. It is a non-specific agent able to hydrolyzenecrotic tissues. This property is due to the presence of sodiumhydroxide (NaOH). The tissue dissolving level (e.g. mainly necrotictissues) is in accordance with NaOCl concentration, contact surface(Hand et al., 1978), contact time and NaOCl solution amount used (The etal., 1979).

[0010] Thus, even if a NaOCl concentration lower than 0,5% is not goodenough to totally dissolve necrotic tissues, the reduced toxicity ofthese low concentrations is interesting. However, this decreased abilityto dissolve necrotic tissues may be made up for by a NaOCl temperatureincreased to 37° C., even if at this temperature, the NaOCl stability isbelow 24 hours.

[0011] b. HOCl and taurine N-monochloramine stability in aqueoussolution.

[0012] Sodium hypochlorite (NaOCl):

[0013] Sodium hypochlorite is a highly unstable molecule. At levelsbelow 5 g/l of available chlorine, its stability is under 2 weeks anddepends on the following factors:

[0014] Light: Sodium hypochlorite is highly sensitive to light andshould be protected by suitable packaging.

[0015] Temperature: NaOCl is highly sensitive to temperature greaterthan 30° C.

[0016] Presence of metal or organic matter: hypochlorite aqueoussolution (containing HOCl molecules) (i.e.: NaOCl+H₂O

HOCl+NaOH) is neutralized by organic matter. Hypochlorite solution isefficient both when it can act readily and when it is in excess incomparison to an organic matter amount.

[0017] pH value: EP 0471129 A1 has established that a pH value between10 and 10.5 yields a high stability to NaOCl oxidative activity (greaterthan 24 months).

[0018] Taurine N-chloramine:

[0019] At a physiological pH (7.4) and at 37° C., the taurineN-chloramine is the more stable of the chloramines (the oxidativeactivity decrease is below 5%/hour at 37° C.) (Grisham M B, Jefferson MM, Melton D F, Thomas E L—J.Biol. Chem. 1984; 259: 10404-13). However,in aqueous solution, the solubility of taurine N-chloramine sodium saltwith a pH value between 7 and 8 is greater, but has a lower stability ofits oxidative activities (DE 4041703 A1), and at a pH=8.3, the stabilitydecreases by around 30% in 15 days followed by a decrease of around0.71% per day (i.e. this equals a decrease of around 61% in 65 days).

[0020] c. Cell toxicity and viability.

[0021] Cell toxicity results mainly from an intracellular protein loss,which generates both an adherence decrease to substrates and celldeformation.

[0022] Cell viability alteration results mainly from the irreversibledecrease of mitochondrial activity and therefore, a reduction of energygenerated by cell respiratory.

[0023] The vulnerability of different cell organisms to NaOCl and TauCldepends on many factors:

[0024] The exposition level of the cell surface. Thus, cell systems witha high cell organization e.g. in epithelium and dental plaque are lesssensitive (i.e. surface cells are sacrificed for profound cells) thanone-cell systems (prokaryotes, mammalian mobile cells, or other one-cellsystems).

[0025] Membrane type that protects intracellular elements (i.e. membranepermeability level to oxidants). The most efficient are viral proteinicmembranes.

[0026] A membrane presence that protects key intracellular systems(e.g., DNA (nucleus), energetic production (mitochondria), secretionprocess (Golgi's apparatus), etc.). Prokaryotes do not possess theseprotector systems and, consequently, are more vulnerable.

[0027] The intracellular antioxidant amount (i.e., gluthatione, acetylN-cysteine, taurine, amino acids, thiol groups, etc.) that is specificfor each cell type. Prokaryotes possess a down antioxidant level.

[0028] The extracellular antioxidant amount (i.e. taurine, thiol groups,organic matter, metal, blood, extracellular matrix, etc.).

[0029] The liquid flux level that irrigates cells and, consequently,dilutes oxidants.

[0030] The exposition time to oxidants.

[0031] The local physicochemical environment (e.g. surface-active,oxidants, olfactory or gustatory properties, pH, pKa, density,solubility, viscosity, coloration, water-ectanol sharing factor).

[0032] In a therapeutic treatment in vivo, the factors described aboveshould be integrated for the determination of active agent levels toadapt them to both clinic status and therapeutic aims.

[0033] i) Sodium hypochlorite (NaOCl) or hypochlorous acid (HOCl):

[0034] On the rat macrophage like-cells RAW 264.7, with a (NaOCl)=1 mM(NaOCl concentration), the cell viability is highly altered(irreversible)(Park E. et al., 1997).

[0035] On the mouse macrophages, with (HOCl)>0.125 mM, cell deathincreases significantly. This toxicity is abolished by a nitrite (NO₂ ⁻)excess (NO₂ ⁻ alone does not generate cytotoxic activity) (MarcinkiewiczJ. et al., 2000).

[0036] On human macrophages, fibroblasts and keratinocytes, in vitro:

[0037] With (NaOCl)=13.433 mM, toxicity is so great that it cannot beneutralized by antioxidants (i.e. with physiological concentrations).

[0038] With (NaOCl)>6.7165 mM, NaOCl has a high toxicity.

[0039] With (NaOCl)<3.358 mM, toxicity can be neutralized by anantioxidant addition.

[0040] With (NaOCl)<1.679 mM, toxicity is very low with an antioxidantpresence (Hidalgo E. & Dominguez C., 2000).

[0041] The adherence loss of macrophages generated by HOCl: With(NaOCl)=1.0075 mM, after two hours of contact in vitro, 95% of the cellsare alive but only 40% keep their adherence to substrates.

[0042] On human endothelial cells in vitro (Pullar J M et al., 1999):

[0043] With [HOCl]≦25 μM, HOCl is not toxic.

[0044] With [HOCl]>25 μM, cell toxicity increases progressively(exposition time-dependent).

[0045] With [HOCl]=50 μM, some cell contractions were observed, thecells became rounded within the first 10 minutes and some lost theiradherence after one hour and the majority after three hours.

[0046] On human fibroblasts in vitro:

[0047] With (NaOCl)≧1,0075 mM (observed for 24 hours after a 15-minuteexposition) cell viability is altered.

[0048] With (NaOCl)=16,791 mmol/l cell morbidity is complete.

[0049] For 67,165 μmol/l<(NaOCl)<671,655 μmol/l, 100% of cells arealive.

[0050] With (NaOCl)<671,655 μmol/l, and a FCS presence (2%), cellviability is not altered (24 hours of exposure) and both growth and cellproliferation are stimulated (the latter enhance with the (NaOCl)decrease and with a highest efficiency at 33,582 μmol/l) (Hidalgo E. &Dominguez C., Life Sci. 2000 August 4;67(11):1331-44).

[0051] With (HOCl)<50 μM, HOCl does not alter in vitro human fibroblastskin viability and does not induce cell apoptose (Vile G. F. et al.,2000).

[0052] ii) The effects of taurine N-chloramine (TauCl) on cellviability:

[0053] On rat C6 glioma cells, a (TauCl)=0˜2 mM does not alter cellviability in vitro (Liu Y. et al., 1999).

[0054] On human skin fibroblasts, a (TauCl)≦100 μM does not inducecytotoxicity or cell apoptose in vitro (Vile G. F. et al., 2000).

[0055] On human synoviocytes-like fibroblastes, with (TauCl)=400-500 μM,cell morphology changes (˜30%-50% of cells took a rounded form and losttheir adherence to the plastic surfaces) although viability has beenpreserved (>95%) (Kontny E. et al., 1999).

[0056] On mouse T cells:

[0057] With (TauCl)=30-300 μM, cell viability is not altered (i.e.mitochondrial activity).

[0058] At 300 μM, TauCl is cytotoxic (Marcinkiewicz J. et al., 1998).

[0059] On mouse dendritic cells incubated 24 hours with TauCl:

[0060] For 0.05 mM<(TauCl)<0.5 mM, mitochondrial activity (cellviability) is not altered.

[0061] With (TauCl)>0,5 mM, cell viability decrease significantly(Marcinkiewicz J. et al., 1999).

[0062] On macrophages or macrophage-line cells, with a (TauCl)=50˜600μM, cell viability is not altered. (TauCl)>1 mM alters it (MarcinkiewiczJ. et al., 1995).

[0063] d. Cellular take-up of exogenous HOCl and taurine N-chloramine.

[0064] HOCl is a lipophilic oxidant and, consequently, easily andreadily cross cell membranes (i.e. ˜80% of HOCl molecules are taken upby human fibroblasts within the first 10 minutes) (Vile G. F. et al.,2000). In vitro with (HOCl)=35 μM, endothelial cells take up 50% of HOClmolecules within ½ minute and 100% within 15 minutes, with a highmajority within the first 10 minutes (Pullar J. M. et al., Am J Physiol.1999 October; 277(4 Pt 2): H1505-12).

[0065] TauCl is taken up by specific transport systems. Therefore, invitro, the K_(m) and the V_(max) values in relaxed rat RAW264.7 cellsare 23.3 μM and 51.3 pmol/min/10⁶ cells, respectively (K_(m)=28,1 μM andV_(max)=90,9 pmol/min/10⁶ cells for taurine).

[0066] In LPS-stimulated macrophages, K_(m)=45,9 μM and V_(max)=82,6pmol/min/10⁶ cells for TauCl, and K_(m)=17,3 μM and V_(max)=116,3pmol/min/10⁶ cells for taurine.

[0067] Membrane transport systems are specific to each of thesemolecules and depend on Na⁺ level, temperature, and energy.

[0068] The blood biodistribution of TauCl and taurine induce a readytake up by cells of liver, lung, spleen, stomach, intestine and kidneys.In addition, cells present within an inflammatory site readily take upthese two molecules (with a inflammation/blood ratio equal to 6.43 and4.84 respectively) (Kim C. et al., 1998). Others data show a ready takeup by kidneys, liver, spleen, and marrow. The take up by heart andmuscle is slow (Huxtable R J, J. Nutr. 1981; 111:1275-86).

[0069] e. Antiseptic properties.

[0070] Sodium hypochlorite is a very strong and efficient bactericidal,virucidal and fungicidal agent (Shih et al., 1970; Bloomfield & Miles,1979, Harrison & Hand, 1980). The bactericidal minimum concentration ofNaOCl (i.e. for Gram−and Gram+bacteria) is 3.36 mM (0.025%) (Heggers J.P. et al., 1991) and the minimum virucidal concentration for VIH is19.062 mM (1%) of available chlorine.

[0071] In contrast, TauCl has a very low bactericidal activity. Onlydichloramines generate some bactericidal activity (i.e. with E. Coli inacidic conditions) (Marcinkiewicz J. et al., 2000).

[0072] 3. Inflammation.

[0073] Inflammation is a defense mechanism toward all aggression types.Sentinel cells (e.g. macrophages and dendritic cells (DC)), thatgenerate an immune system initialization via both a generation and arelease of mediators detect an aggressor (Marcinkiewicz J. et al.,1999). These mediators induce a reaction cascade and both activate andregulate the immune system in an adaptive manner to the aggression type.After the aggressor agents are removed, a regulatory system generates aninflammation turnover followed by a healing/regeneration process.

[0074] Two immunity types are perceived: innate (natural) and acquired(adaptive).

[0075] The cell part of the innate (natural) immunity is made up ofmonocytes (mononuclear phagocytes), polymorphonuclear neutrophils (PMN),and natural killer cells (NK). These cells use the complement cascade,or some recognition protein, e.g., reactive protein C and amyloidprotein. These proteins are able to attach themselves to carbohydratemolecules present on bacteria membranes. PMNs are included in the firstmammalian defense line and cooperate closely with macrophages (one ofthe major effector cells of the immune system). PMNs are responsible forthe non-specific defense in acute inflammation and macrophages take asimilar role in both acute and chronic inflammations (Marcinkiewicz J.et al., 1994).

[0076] The acquired (adaptive) immunity involves several T cell typesand uses antibodies as effector proteins. T cell receptors andantibodies are recognition molecules. B cells recognize carbohydrates,proteins, and some simple chemical structures while T cells recognizeonly peptides.

[0077] Dendritic cells (DC) play an important role. Under inflammatorymediator action, DCs migrate from non-lymphoid tissues to lymphoidorgans where they lose their ability to scavenge antigens and acquire anincreasing ability to stimulate T cells (Marcinkiewicz J. et al., 1994).

[0078] 4. Inflammatory mediators.

[0079] Cytokines are the most important intercellular messengermolecules of the immune system (Megarbane B. et al., 1998). Cytokinesare generated and released from activated immune cells and they inducesome particular biological activities after binding to a specific targetcell receptor, in an autocrine or a paracrine manner. Macrophages and Tcells are main productive cells of cytokines, although many other cellsalso can produce them. Cytokines are main and real regulators of bothhumoral and cellular immune response. Cytokines travel together and thebalance of their activities is crucial for immune system regulation,e.g., via a competition between TH1 (IL-2, INF-γ, TNF-βand IL-12) andTH2 (IL-4, IL-5, IL-10 and IL-13) T cells.

[0080] TH1 cells are involved in cell immunity and are responsible forcytotoxic activities of macrophage, T cells and natural killer cells.

[0081] TH2 cells are associated with humoral response, and, for example,IL-10 (i.e. a TH2 type cytokine) strongly inhibits effective functionsof macrophages and TH1 cells (Marcinkiewicz J., 1997).

[0082] Cytokine regulatory functions can be extended to a selection ofimmunoglobulin isotypes during humoral response. Thus, selectiveinhibitions of cytokines generate an immune response modulation.

[0083] Eicosanoids (prostaglandins and leukotrienes) and nitric oxide(NO), produced by activated macrophages, have an important role in theregulation of cytokine production. Eicosanoids are generated fromarachidonic acid, which is derived from cell membrane phospholipides.

[0084] Prostaglandines (PG) are generated under the cyclooxygenase (COX)catalyzing action. Two cyclooxygenase types are distinguished: theconstitutive form (COX1) and the induced form (COX2). COX2 production isactivated within inflammatory cells by pro-inflammatory mediators. Thus,COX2 catalyzes the synthesis of prostaglandins E₂ (PGE₂) andprostacyclins I₂ (PGI₂) in macrophages, and prostaglandines D₂ in mastcells.

[0085] Prostaglandins (particularly PGE₂) and leukotrienes (particularlyLTB₄) change immune responses. Therefore, equilibrium in both productionand effects of these eicosanoids is needed to induce a harmoniousfunctioning of the immune system.

[0086] Nitric oxide (NO) is synthesized from L-arginine under thecatalyzing action of the constitutive nitric oxide synthetase ((cNOS)that is calcium dependent) or the induced nitric oxide synthetase((iNOS) that is calcium independent).

[0087] cNOS permits the synthesis of the basic form of nitric oxide (NO)in cells of both endothelium and nervous system.

[0088] iNOS is found in a variety of cells including macrophages,neutrophils and hepatocytes. NO generation plays an important role inmacrophage cytotoxicity and their ability to kill pathogenmicroorganisms and, consequently, in mammalian non-specific defenseagainst many pathogens and tumor cells.

[0089] More characteristics of these inflammatory mediators aredescribed in Knight JA et al., 2000; Marcinkiewicz J. et al., 1997; andMegarbane B et al., 1998.

[0090] 5. The influence of hypochlorous acid and taurine N-chloramine onan inflammatory site.

[0091] On Bacteria.

[0092] Rat peritoneal macrophages stimulated by non-chlorinatedGram+bacteria (Staphylococcus aureus, S. epidermidis, and Escherichiacoli) release high concentrations of nitric oxide, TNF-α, and IL-6. Thesame bacteria chlorinated by HOCl lose their abilities to induce anitric oxide and TNF-α release while IL-6 production and phagocytosisare not altered (Marcinkiewicz J. et al., 1994).

[0093] On Endothelium.

[0094] HOCl increases the endothelium permeability and promote leukocyteadherence to microcirculation endothelium. Taurine N-chloramine reducesan endothelium permeability increase generated by PMN activities.Taurine alone is without effect (Tatsumi & Flies, 1994).

[0095] On Cellular Growth.

[0096] In vitro, on endothelial cells of the human umbilical vein, aHOCl down level (5 nM/1.2×10⁵ cells) does not induce a cell death but atemporary stop of cell growth (Vissers M C et al., 1999). In addition,low concentrations of both HOCl and physiological chloramines lead invitro to an inhibition of DNA synthesis and cell division on skinfibroblasts (Vile G F et al., 2000).

[0097] On Non-free Proteins (e.g. Collagen, etc.).

[0098] HOCl is a very strong oxidant. In addition, HOCl chlorinatesproteins and makes them more vulnerable to an endopeptidase-degradation.Thus, HOCl contributes to a destruction of the tissue surrounding theinflammatory site. TauCl is an oxidant with lower strength and seems tohave a lesser responsibility for damage to these tissues.

[0099] On Collagenases.

[0100] TauCl induces a direct inhibition/inactivation of collagenaseswhile it has no effect on the collagen proteolytic susceptibility. Incomparison, leucine and alanine N-monochloramines have no inhibitoryeffect on collagenases and increase the proteolytic susceptibility ofcollagen (Davies JMS et al., 1994).

[0101] On Free Proteins (Ovalbumin, Bacterial Enzymes, etc.).

[0102] Free protein chlorination enhances their immune sensitivity,likely via an improvement of both their treatment and presentation byantigen-presenting cells (i.e. macrophages and dendritic cells). Thischlorination is ten times more important for HOCl thantaurine-N-monochloramines (TauCl) but, in vivo, TauCl is more stableand, consequently, TauCl can be regarded as the main physiologicalchlorinating agent (Marcinkiewicz J. et al., 1999).

[0103] On Dendritic Cells (DC) (Marcinkiewicz J. et al., 1999).

[0104] Two hours pre-incubated rat DCs with TauCl underwent aconcentration-dependent inhibitory activity. Thus, a TauCl concentrationequal to 500 μM ((TauCl)=500 μM) almost completely inhibits the DCrelease of reactive oxygen agents (ROS) generated via a respiratoryburst, nitric oxide, PGE₂, TNF-α, IL-6, IL-10, and IL-12. In addition,the lipopolyssacharide-induced expression of MHC type II and moleculeB7-2 is also inhibited. At this concentration, TauCl may be toxic to DCwhen they are exposed for a long time. With (TauCl)=250 μM, TauCl has amore selective action. Therefore, it inhibits the production of IL-10,IL-12, PGE₂, and nitric oxide. TNF-α and ROS generation is notinhibited. In addition, a DC exposition to TauCl seems to promote a TH1response and decreases the TH2 activity.

[0105] On T Cells.

[0106] TauCl inhibits the release of IL-2 and IL-6 by T cellspre-incubated with a (TauCl)=100-300 μM and stimulated with either amitogen, an antigen or an ovalbumin-APC complex (Marcinkiewicz J. etal., 1998).

[0107] On Phagocytes.

[0108] Antigens chlorinated by HOCl or TauCl do not induce an productionof inflammatory mediators by the phagocytes that phagocytosed theseantigens (Marcinkiewicz J. et al., 1994 & 1997).

[0109] On Macrophages.

[0110] Chloramines such as taurine N-mono and (N,N)-dichloramine,N-monochloro-ethanolamine and N-dichlorophosphoethanolamine as well asNaOCl (sodium hypochlorite), all inhibited the release of nitric oxidein a dose-dependent manner. Serine N-chloramine (SerCl) had a lesserhalf-life than TauCl (immediately after its preparation, (SerCl)=300 μMinhibited the nitric oxide generation for 85%; after 24 hours, thisinhibition was reduced to 22%). TauCl inhibited the oxide nitricgeneration for 98% with (TauCl)=600 μM and 8-22% with (TauCl)=100 μM(i.e., this value changes with cell type). This inhibitory effect wasexecuted within the iNOS gene transcription. Taurine alone was withouteffect (Marcinkiewicz J. et al., 1995). HOCl (likely via TauCl activity)and TauCl inhibited COX2 post-transcriptional expression i.e. four-hoursdelay on the kinetic expression of mRNA (and consequently the PGE₂production) and TNF-α transcriptional velocity (i.e., in adose-dependent manner with an IC₅₀=400 μM)(Quinn M R et al., 1996).TauCl inhibits COX2 expression either in non-stimulated andINF-γ-stimulated macrophages. In contrast, in INF-γ-stimulatedmacrophages TauCl inhibits both the iNOS expression and the productionof TNF-α and IL-6. TauCl had no effect on IL-1α production for allstimulation levels. The native taurine alone had no effect on cytokineproduction. In addition, HOCl-oxidized plasma lipoproteins had anability to reduce iNOS mRNA synthesis and, thus, to inhibit the nitricoxide production and contribute to atherosclerotic lesion development(Moeslinger T et al., 2000).

[0111] On Polymorphonuclear Neutrophils.

[0112] TauCl inhibits production of nitric oxide, PGE₂, IL-6 and TNF-αin a dose-dependent manner. Native taurine has no effect. Someexperiments (Marcinkiewicz J et al., 1998 & 2000) with luminolchemiluminescence-dependent (LCL) measures have shown the following:

[0113] Both taurine and TauCl reduced ROS production. However, only hightaurine concentrations altered LCL and taurine activity is lower thanTauCl.

[0114] HOCl reduces myeloperoxidase activity in a retroactivedose-dependent manner. In vitro, TauCl and HOCl inhibit myeloperoxidaseextracted from neutrophils.

[0115] HOCl (250 μM) inhibits hydrogen peroxide production in adose-dependent manner. Taurine (500 μM) or nitrite (250 μM) neutralizesthis inhibition. TauCl has no effect on this production.

[0116] HOCl and TauCl induce a chemiluminescence dose-dependentdecrease, TauCl (IC₅₀=550 μM) is less efficient than HOCl (IC₅₀=100 μM).

[0117] TauCl and taurine inhibit superoxide anion (O₂ ⁻) production bystimulated neutrophils. This inhibition involves a different mechanismthan those implicated in TauCl formation (i.e., association of thetaurine (or TauCl) with a myeloperoxydase specific inhibitor generates asynergic effect).

[0118] However, high concentrations of taurine alter LCL. This activityis less important than TauCl (Marcinkiewicz J et al., 1998).

[0119] On Polymorphonuclear Eosinophils.

[0120] HOCl inactivates sulfidopeptide LTC4 sulfoxides and 6-trans-LTB4leukotrienes only in an extracellular environment (Owen W F et al.,1987).

[0121] On Rat Glioma Cells C6.

[0122] In the central nervous system of activated glioma cells, TauClinhibits production of monocyte chemoattractant protein-1 (MCP-1) andmacrophage inflammatory protein-2 (MIP-2) both in dose-dependent andpost-transcriptional manners (Liu Y et al., 1999). In addition, TauClinhibits both the iNOS gene transcriptional expression (i.e., nitricoxide production) and the COX2 expression (i.e., PGE₂ production) via apost-transcriptional mechanism (Liu Y et al., 1998).

[0123] On Fibroblasts.

[0124] In rheumatoid arthritis patients, TauCl inhibits fibroblast-likesynoviocyte proliferation and decreases the activity of majortranscriptional factors of both IL-6 (IC₅₀˜225 μM) and IL-8 (IC₅₀˜450μM) in a dose-dependent manner. Thus, TauCl reduces both IL-6proinflammatory action and immune cell ability to migrate within aninflammatory site (via an IL-8 inhibition). Whereas IL-6 inhibition isindependent of the fibroblast stimulating agent used (e.g. TNF-α, IL-1βor IL-17), IL-8 inhibition is dependent on the stimulation via TNF-α orIL-1β, but not via IL-17. This shows different signaling pathways fromTNF-α/IL-1β and IL-17 triggered-transduction (Kontny E et al., 1999).These signaling pathways are dependent on two transcription factors:NF-κB and AP-1. In addition, TauCl inhibits both spontaneous andbFGF-stimulated syno-viocyte proliferation (Kontny E et al., 2000).

[0125] Low levels of both HOCl and physiological chloramines (NH₂Cl,TauCl and N-chlorinated α-amino acid) inhibit both DNA synthesis andcell division of cultured human skin fibroblasts (Vile G L et al.,2000).

[0126] On Transcription Factors NF-κB and AP-1.

[0127] NF-κB-dependent gene expression may be altered by TauCl activity.In IL-1β-stimulated human synoviocytes, transduction TauCl-inhibition ofIL-6 and IL-8 is executed via a DNA-bonding ability reduction of NF-κBand AP-1. IL-6 transcription is under a NF-κB control, while both NF-κBand AP-1 control IL-8 transcription. Thus, a (TauCl)=250 μM selectivelyreduces the DNA-bonding of NF-κB (i.e., the IL-6 transcription) withoutaltering AP-1 DNA-bonding (i.e., the IL-8 transcription). TauCl acts onboth NF-κB and AP-1 transcription factors to inhibit the IL-6 and IL-8transduction. At 500 μM, TauCl decreases the DNA-bonding activity ofboth NF-κB and AP-1 (i.e., the transcription of IL-6 and IL-8 isreduced)(Kontny E et al., 2000). These two transcription factors areregulated via a redox mechanism ((Sen C. K., Packer L., Fased J. 1996;10:709-20), (Li N. & Karin M., Fased J. 1999; 13:1137-43), (Kunsch C. &Medford R. M., Circ Res. 1999 October 15; 85(8):753-66.)). It seems thatTauCl may interfere the intracellular redox status of thesetranscription factors and, therefore, some anti-inflammatory propertiesmay be suggested from TauCl (Kontny E et al., 2000).

[0128] On Complement.

[0129] The C₅ component of the human complement may be activated byoxidants, e.g., hydroxyl radicals, hypochlorite or chloramines (i.e.,TauCl and mainly NH₂Cl). This activation is due to a C₅ structuralchange induced by a Met. residue oxidation within the C₅ protein withoutpeptide cleavage. These changes lead to a C₆ bonding site expression,which normally is formed after a C₅ specific cleavage in C_(5a) andC_(5b), via one of two C₃/C₅ convertases. The C₅-oxidation product issimilar to C_(5B). Thus, it is able to initiate the combination of theC₅₋₉ membrano-lytic complex.

[0130] Chemotactic fragments are not directly generated, but activatedC₅ components (like C_(5b)) are readily attacked by enzymes such askallikrein, which produce C_(5a)-like fragments that have a chemotacticactivity. It is likely that the C₅₆₇ complex generated with C₅ also havea chemotactic activity (i.e., similarly to C_(5b67) complex). Inaddition, the C_(5b-9) complex is known to stimulate PMNs at non-toxicconcentrations. Thus, the same property may be suggested for thecorresponding C₅₋₉ complex and, consequently, this may lead to a viciouscircle that increases tissue lesions (Vogt W, 1996).

SUMMARY OF THE INVENTION

[0131] This invention relates to a pharmaceutical composition includingat least one halogenated compound, and at least one N-halogenatedderivative of at least one compound selected from zwitterionic and/oramino acid compounds, where the composition does not generatesubstantial stimulation of myeloperoxidase activity in a mammal.

[0132] This invention also relates to a method of preparing apharmaceutical composition including mixing at least one halogenatedcompound and at least one zwitterionic compound and/or at least oneamino acid or their derivatives, and optionally at least one excipientto obtain at least one N-halogenated derivative, and at least onehalogenated compound in a sufficient therapeutic amount to notsubstantially stimulate myeloperoxidase activity in a mammal.

[0133] This invention further relates to a method for treatment and/orpreventing viral infections, and/or bacterial infections, and/orparasitical infections and/or fungal infections and/or diseasesgenerated from non-conventional transmissible agents, in humans oranimals including administering to a human or animal a pharmaceuticallyeffective amount of a pharmaceutical composition including at least onehalogenated compound, and at least one N-halogenated derivative of atleast one compound selected from zwitterionic compounds and/or the aminoacids or their derivatives without substantial stimulation ormyeloperoxidase activity in the human or animal.

DETAILED DESCRIPTION

[0134] I have discovered that in inflammatory sites, beyond anybactericidal activity, NaOCl contributes to (1) an increase in thetransition to the cleansing of necrotic and suppurating mass, (2)stimulates local immunity and (3) activates the tissue regenerationprocess. These abilities are induced from sodium hypochlorite (i.e.,hypochlorous acid (HOCl) properties and the hydrolysis generated fromsodium hydroxide (NaOH)) and its N-chlorinated derivatives.

[0135] Consequently, this invention provides a pharmaceuticalcomposition comprising (i) at least one halogenated compound and (ii) atleast one N-halogenated derivative of at least one compound selectedfrom zwitterionic and/or amino acid compounds.

[0136] Within compositions according to the invention, the halogenatedcompound (i) is an antiseptic.

[0137] Amino acids included in the constitution of compositionsaccording to the invention can be natural amino acids, derivatives oranalogous of the latter.

[0138] More particularly, the halogen of the (i) halogenated compoundsand the (ii) N-halogenated derivatives of the invention composition,similar or different, may be fluorine, iodine, bromine, and mainlychlorine.

[0139] Favorably, the halogenated compound (i) is an alkaline metalhypochlorite, and preferably the sodium hypochlorite, and theN-halogenated derivative (ii) is an N-halogen derivative of taurine andpreferably a taurine N-halo-amine and even more preferably taurineN-chloramine.

[0140] The invention composition is remarkable from its robustproperties such as large spectrum of application such asanti-inflammatory, immunity modulation, and tissue healing stimulationas well as those without stimulation of myeloperoxidase activity.

[0141] The hypochlorite titer of the invention composition is preferablybelow or equal to about 1 mole/liter of available chlorine, and can beadapted to clinical use. Usefully, the invention composition contains ahypochlorite of alkaline metal. Preferably, the invention compositioncontains a sodium hypochlorite q.s. with a minimum titer of availablechlorine that is greater than or equal to about 1 picomole/liter.

[0142] The N-chloramine titer of the invention composition is preferablyless than or equal to abut 5 moles/liter, and may be adapted to clinicaluse. Usefully, the invention composition contains an N-halogenatedderivative, such as the taurine N-chloramine, with a concentrationbetween about 5 moles/liter and about 0.01 femtomoles/liter. Preferably,the invention composition contains a N-halogenated derivative such asthe taurine N-chloramine, q.s. with a minimum titer greater than orequal to about 0.01 femtomoles/liter.

[0143] The (i) halogenated compound and the (ii) N-halogenatedderivative are associated in the composition according to the inventionwith an excipient, such as purified water, in accordance withtherapeutic use. Preferably, it concerns an osmotic (isotonic) purifiedwater. This excipient may contain diverse agents, pharmaceuticallycompatible with both (i) the halogenated compound and (ii) theN-halogenated derivative, and which can allow for modification of somephysicochemical properties such as stability, pH, pKa, density,solubility, viscosity, coloring, water/ectanol sharing factor, andsurface-active, oxidative, olfactory, or gustatory properties of theinvention composition via a suitable agent addition. The inventioncomposition may also contain some anti-oxidants and/or amino acids thathave a dilution effect via neutralization of some alkaline metalhypochlorite molecules. These anti-oxidants, amino acids and theirN-halogenated derivatives should have a neutral pharmacological activityor its activity should be pointed to therapeutic aims and should notexercise a direct stimulation of myeloperoxidase activity in thepresence of invention composition active agents.

[0144] The invention also concerns the preparation of the compositiondescribed above. Thus, this composition can be sold in a form to preparebefore use, i.e., (i) the halogenated compound(s) can be mixed with (ii)the N-halogenated derivative(s) and one or several excipients. Thispresentation form can be considered if it is required to guarantee thebest time stability of the composition and, in particular, the activeagents that constitute the latter. However, even in a presentation wherethe constituting products would be associated, the invention compositioncan be sold with an excipient, such as purified water according to thetherapeutic use. Preferably, this should be an osmotic (isotonic)purified water. In addition, this excipient may contain diverse agentspharmaceutically compatible with the totality of final compositionmolecules, which allow for the modification of some physicochemicalproperties of the invention composition via an addition of suitableagent(s) such as stability, pH, pKa, density, solubility, viscosity,coloring, water/ectanol sharing factor, and surface-active, oxidative,olfactory, or gustatory properties.

[0145] The invention composition can also be prepared before itsadministering to the patient via a mixture comprising:

[0146] (i) at least one halogenated compound, and

[0147] (ii) at least one N-halogenated derivative of at least onecompound selected from zwitterionic and/or amino acid compounds, andtheir derivatives.

[0148] More particularly, the halogen(s) of the halogenated compound (i)and the N-halogenated derivative (ii) may be selected from fluorine,iodine, bromine, and/or chlorine, most preferably chlorine.

[0149] Favorably, the halogenated compound (i) is a halide such as analkaline metal hypochlorite, and preferably the sodium hypochlorite, andthe N-halogenated derivative (ii) is a taurine N-halogenated derivativeand preferably a taurine N-haloamine and even more preferably thetaurine N-chloramine.

[0150] The aforementioned halogenated compound(s) (i) are usefullydisplayed in a liquid or semi-liquid (such as a gel) solution form,favorably within an excipient as described below. These solutions,advantageously hypochlorite solutions, may be stabilized in accordancewith the patent EP 0 471 129 A1 via a pH regulatory agent to generate apH between 10 and 10.5 with respect to cell viability.

[0151] The aforementioned N-halogenated derivative(s) (ii) are usefullydisplayed in a liquid or a semi-liquid (such as a gel) solution form,favorably within an excipient as described below.

[0152] Favorably, the invention composition may be prepared via amixture of the two solutions described above with at least one excipientaccording to therapeutic use such as purified water. It preferablycontains the osmotic (isotonic) purified water. In addition, thisexcipient can contain diverse agents, pharmaceutically compatible withall molecules of the final mixing to modify some physicochemicalproperties of the invention composition such as stability, pH, pKa,density, solubility, viscosity, coloring, water/ectanol sharing factor,and surface-active, oxidative, olfactory, or gustatory properties via anaddition of suitable agent(s).

[0153] In addition to the process described above, the inventioncomposition may be prepared via a mixture of the two followingsolutions:

[0154] (i) at least one halogenated compound as defined above, which isusefully displayed in a liquid or a semi-liquid (such as a gel) solutionform, preferably within an excipient as described above,

[0155] (iii) at least one zwitterionic compound and/or at least oneamino acid and/or at least one primary or secondary amine, (thezwitterionic compound and/or amino acid and/or primary or secondaryamino amine are later referred to as “Zw/Aam”), which is usefullydisplayed in a liquid or a semi-liquid (such as a gel) solution form,favorably within an excipient as described above, to obtain anassociation of both (i) at least one halogenated compound and (ii) atleast one N-halogenated derivative, and this with a sufficienttherapeutic amount of molecules to inhibit myeloperoxidase activity.

[0156] This mixture is preferably realized with an excipient as definedabove.

[0157] In case Zw/Aam is an amino acid, it preferably concerns taurineor a taurine pharmaceutical analog.

[0158] In this realization method, when the antiseptic halogenatedcompound (i) is a halide such as alkaline metal hypochlorite (which isan alkaline metal salt of hypochlorous acid), derivatives generated willbe N-chlorinated, and these will more particularly be N-chloramines.

[0159] The hypochlorite titer of the first active solution (i) shouldtake into consideration the stoichimetry and reactivity level of thereaction between hypochlorous acid and Zw/Aam molecules. In case thisreaction is not complete, remaining Zw/Aam molecules should notstimulate myeloperoxidase activity in the presence of inventioncomposition active agents.

[0160] In case the stoichimetry is 1/1 and with a complete reaction(e.g., between hypochlorous acid and taurine), the hypochlorite titer ofthe first active solution is preferably lower than or equal to about 6moles/liter of available chlorine, and must be adapted both to theZw/Aam molecule amount of the second solution and to clinical status. Inthis preparation method, the halide solution (i) favorably contains analkaline metal hypochlorite. Even more preferably, the haloid solution(i) contains sodium hypochlorite q.s. with an available chlorine titerbetween abut 6 moles/liter and about1,000.01 femtomoles/liter. Thetaurine titer of the second solution (iii) of this invention preparationmethod is preferably lower than or equal to about 1 moles/liter and maybe adapted to clinical use. It is useful for the second solution (iii)of this invention preparation method to have a taurine concentrationbetween about 5 moles/liter and about 0.01 femtomole/liter. Even morepreferably, the second solution (iii) of this preparation method has ataurine titer greater than or equal to about 0.01 femtomole/liter.

[0161] The excipient(s) preferably added in methods described above maybe used as a secondary diluting solution with the aim to adapt thetreatment to the clinical status. It usefully concerns osmotic(isotonic) purified water. This excipient will favorably be similar tothe excipient used for the compounds and derivatives that have beenmixed, and if they are not identical, the excipient should bepharmaceutically compatible to be mixed with the other excipient(s),before all clinical uses. In addition, this excipient can containdiverse agents, pharmaceutically compatible with all molecules of thefinal therapeutic mixture with the object of modifying somephysicochemical properties of the invention composition such asstability, pH, pKa, density, solubility, viscosity, coloring,water/ectanol sharing factor, and surface-active, oxidative, olfactory,or gustatory properties via an addition of a suitable agent(s).

[0162] This excipient may contain anti-oxidants and/or amino acids thatwill have both a dilution effect and an oxidant neutralization of theactive solution (i) (e.g., the alkaline metal hypochlorite). Theseanti-oxidants, amino acids and their halogenated derivatives should havea neutral pharmaceutical activity or a pharmaceutical activity inducingthe desired therapeutic effect. In all cases they should be both lesstoxic than the oxidants of (i) the main active solution andpharmaceutically compatible with all molecules of the final therapeuticsolution.

[0163] The composition according to the invention can also be sold in aform adapted to local use, e.g., a gel or an aerosol.

[0164] The above-mentioned invention composition is particularly usefulin humans or animals for treatments of viral infections and/or bacterialinfections and/or parasitical infections and/or fungal infections and/ordiseases generated from non-conventional transmissible agents; and/orfor treatments of chronic, progressive or acute inflammation; and/or forimmunity modulator treatments; and/or for tissue regeneration stimulatortreatments. In addition, the therapeutic composition may be used inpre-surgical irrigations and/or per-surgical irrigations and/orpost-surgical irrigations.

[0165] The invention concerns particularly the local treatment ofinfections due to herpesviridiae family virus.

[0166] The invention composition is preferably used locally aiming toremove secondary effects, e.g., atherosclerosis. It can be applied toall external or internal mucous (e.g., oral, genital, vaginal,ophthalmic, otic, sinusal, nose-and-throat, dermal, and the like). Theinvention composition may appear under an adapted form for thisadministration, such as in a semi-liquid form (e.g., a gel) via anaddition of one or several compatible pharmaceutical substances e.g.,cellulose, amino acids, peptides, and/or proteins.

[0167] The invention composition may also be adapted to clinical statusand/or injured mucous. This adaptation is executed via a concentrationchange of active products of the therapeutic solutions.

[0168] For non-restrictive examples of such adapted therapeuticsolutions:

[0169] i) For an infection treatment.

[0170] For endodontic treatment, concentrations between about 1 andabout 0.2 moles/liter of sodium hypochlorite, and approximately betweenabout 100 to about 0.001 picomoles/liter of TauCl are preferred (i.e.,these concentrations vary with organic matter amount present in canals).

[0171] With highly stained keratinized mucous (with profuse presence oforganic matter (infectious agents, blood, profuse and varyingsecretions, suppurating discharge, etc.)), a sodium hypochloriteconcentration between about 0.1 and about 0.02 mole/liter and a TauClconcentration between about 1 and about 0.001 picomoles/liter arepreferable (non-restricting example).

[0172] With moderately stained keratinized mucous (with some organicmatter visible on a compress after a gentle friction, for example), asodium hypochlorite concentration between about 20 and about 10millimoles/liter of available chlorine and a TauCl concentration betweenabout 1 and about 0.01 nanomole/liter are preferable (non-restrictingexample).

[0173] With clean keratinized mucous (without organic matter visible),preferred concentrations may be between about 10 and about 2millimoles/liter of available chlorine for sodium hypochlorite (NaOCl),and between about 50 and about 1 micromoles/liter for TauCl(non-restricting example).

[0174] With highly stained non-keratinized mucous, concentrations may bebetween about 50 and about 10 millimoles/liter of available chlorine forNaOCl and between about 0.1 and about 0.001 picomoles/liter for TauCl(non-restricting example).

[0175] With moderately stained non-keratinized mucous, concentrationsmay be between about 10 and about 5 millimoles/liter of availablechlorine for NaOCl and between about 1 and about 0.01 nanomoles/literfor TauCl (non-restricting example).

[0176] With clean non-keratinized mucous, concentrations may be betweenabout 5 and about 0.8 millimoles/liter of available chlorine for NaOCland approximately between about 50 and about 1 micromoles/liter forTauCl (non-restricting example).

[0177] With important and sensible organs (eyes), concentrations mustboth be the least toxic (via a high dilution or a specific scavenge ofNaOCl by an antioxidant addition) and executed with a profuse dilutedirrigation:

[0178] For stained organs, concentrations may be between about 5 andabout 0.1 millimoles/liter of available chlorine for NaOCl and betweenabout 1 and about 0.01 femtomoles/liter for TauCl (non-restrictingexample).

[0179] For non-stained organs, concentrations may be between about 0.1and about 0.01 millimoles/liter of available chlorine for NaOCl andbetween about 50 and about 1 micromoles/liter for TauCl (non-restrictingexample).

[0180] ii) For the treatment of non-stained organs with the object ofimmune stimulation and/or tissue regeneration, concentrations could bebetween about 500 and about 1 micromoles/liter of available chlorine forNaOCl and between about 200 and about 10 micromoles for TauCl(non-restricting example).

[0181] The composition according to the invention is useful for localtreatment of diseases or inflammatory processes that can be chronic,and/or progressive and/or acute. The composition is also recommended forpre- surgical irrigation and/or per-surgical irrigation and/orpost-surgical irrigation of internal and/or external mucous and ofopened-injures. The invention more particularly concerns a treatmentmethod of lesions and infections described above, which comprisescontacting the invention composition on mucous that must be treated,(for non-restricting example) between 2 and 3 times a day andapproximately during 20 to 60 seconds, not followed by a rinsing. Thecomposition amount employed should be sufficient to not generate a totalneutralization of the therapeutic active agents. In the therapeutic use,the invention solution should not stay static. Concentrations of thecomposition invention should be adapted to the evolution of the clinicalstatus until healing.

[0182] The invention more particularly concerns the local treatment oflesions and infections linked to chronic and/or acute parodontitis.Thus, the invention composition is usefully adapted for irrigation ofperiodontal pockets, with the aim for removing these periodontal pocketsas the composition has both antiseptic and anti-inflammatory activities,and acts as an immunity modulator and healing stimulator of periodontaltissues (i.e., alveolar bone, alveolodental ligament and gingiva).

[0183] Chronic periodontitis is a disease mainly due to pathologicaction of anaerobic bacteria, and particularly Actinobacillusactinomycetemcomitans, Porphyromonas gingivalis, Bacteroides forsythusand Prevotella intermedia. These bacteria induce chronic inflammatoryprocesses that generate a progressive destruction of periodontal tissues(teeth supporting tissue). Periodontitis may result in the removal ofbone tissue followed by tooth loss.

[0184] Whatever the treatment phase of chronic periodontitis,periodontal pocket irrigations have to be executed in the presence of astrong surgical vacuum extraction with the object of avoiding swallowingor inhalation of the therapeutic solution by the patient.

[0185] i) Attack treatment (i.e., between two and three weeks up to thedisappearance of bleeding in probing depth of periodontal pockets).

[0186] J 1: after an assessment of the clinical status, crevicularspaces (with or without periodontal pockets) of oral cavity teeth shouldbe irrigated. A full mouth followed by a tongue brushing, with a mixedsolution of 0.1% chlorhexidine and 0.3% hydrogen peroxide, should beprescribed twice a day (far from the teeth brushing) over ten days, thentwice to three times a week ad vitam aeterman (however, in halitosis,the initial attack treatment should be repeated). Two or threeappointments should be scheduled.

[0187] In the other sessions, the following process will be recommended:education, checking, and motivation for periodontal hygiene; meticulousirrigation (1 ml minimum of the highly stained keratinized mucoussolution for each periodontal site); meticulous scaling and rootplanning.

[0188] When all root surfaces are planned and cleaned, a probing session(this consists first of an irrigation followed by a probing depth)should be executed to evaluate the degree of periodontal disease. Somecomplementary examinations can be made such as sampling picks up andbiological examinations.

[0189] ii) Primary curative treatment (i.e., four weeks).

[0190] Meticulous irrigation of periodontal pockets once every ten daysapplying the solution for moderately stained keratinized mucous, exceptfor sites with a profuse dental plaque (such as interdental furcations)where the solution for highly stained keratinized mucous should beapplied.

[0191] At the last session of the primary curative treatment, anirrigation follows probing depth and root planning.

[0192] iii) Secondary curative treatment (until clinical removal of theperiodontal pockets).

[0193] Meticulous irrigation of periodontal pockets once every ten dayswith the solution for clean keratinized mucous, except for sites with +or − profuse dental plaque (e.g., interdental furcations) where thetherapeutic composition for highly or moderate stained keratinizedmucous should be used.

[0194] Every three sessions of the secondary curative treatment, anirrigation follows probing depth of periodontal pockets and rootplanning.

[0195] iv) Maintenance treatment.

[0196] In any diagnostic of clinical healing, a maintenance treatmentshould be executed. This treatment type is similar to the secondarycurative treatment except that appointments should be made once everythree weeks.

[0197] If after two mouths of treatment a notable healing rather than arecurrence is observed, the last treatment phase—supervision—can beinitiated.

[0198] In case of recurrence, the treatment should be started again at astage that depends on the clinical status observed, i.e., the attacktreatment, or the primary or the secondary curative treatment.

[0199] v) Supervision.

[0200] An appointment should be made once every six weeks. A meticulousprobing depth will be practiced.

[0201] In case of no-recurrence, all crevicular spaces should beirrigated with the solution for moderate stained or clean keratinizedmucous, followed by a meticulous root planning.

[0202] In case of recurrence, the treatment should be started again at astage that depends on the clinical status observed (i.e., the attacktreatment, or the primary or the secondary curative treatment).

[0203] In addition, the invention also concerns bone-filling surgicalperiodontal treatments with some biomaterials associated with theinvention composition and/or one of its components.

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It is claimed: 1) A pharmaceutical composition comprising: (i) at leastone halogenated compound, and (ii) at least one N-halogenated derivativeof at least one compound selected from zwitterionic and/or amino acidcompounds, wherein the composition does not generate substantialstimulation of myeloperoxidase activity in a mammal. 2) A pharmaceuticalcomposition according to claim 1, wherein halogens of (i) thehalogenated compound and (ii) the N-halogenated derivative, which may bethe same or different, are selected from the group consisting offluorine, iodine, bromine and chlorine. 3) A pharmaceutical compositionaccording to claim 1, wherein (i) the halogenated compound is ahypochlorite of an alkaline metal. 4) A pharmaceutical compositionaccording to claim 3, wherein the hypochlorite is sodium hypochlorite.5) A pharmaceutical composition according to claim 1, wherein (ii) theN-halogenated derivative is an N-halogen derivative of taurine. 6) Apharmaceutical composition according to claim 5, wherein the taurine istaurine N-haloamine. 7) A pharmaceutical composition according to claim5, wherein the taurine is taurine N-chloramine. 8) A pharmaceuticalcomposition according to claim 4, wherein the sodium hypochloriteconcentration is between about 1 mole/liter and about 1 picomole/literof available chlorine. 9) A pharmaceutical composition according toclaim 7, wherein the concentration of the taurine N-chloramine isbetween about 5 moles/liter and about 0.01 femtomoles/liter. 10) Apharmaceutical composition according to claim 1, wherein both (i) thehalogenated compound and (ii) the N-halogenated derivative are mixed inwith a pharmaceutically acceptable excipient. 11) A pharmaceuticalcomposition according to claim 1, further comprising a pharmaceuticallycompatible agent which modifies at least one physicochemical property ofthe composition selected from the group consisting of stability, pH,pKa, density, solubility, viscosity, coloring, water/ectanol sharingfactor, and surface-active, oxidative, olfactory, or gustatoryproperties. 12) A method of preparing a pharmaceutical compositioncomprising mixing: (i) at least one halogenated compound, (ii) at leastone N-halogenated derivative of at least one compound selected fromzwitterionic compounds and/or amino acids or their derivatives, and(iii) optionally at least one pharmaceutically acceptable excipient. 13)A method of preparing a pharmaceutical composition comprising mixing: atleast one halogenated compound, and at least one zwitterionic compoundand/or at least one amino acid or their derivatives, and optionally atleast one excipient to obtain at least one N-halogenated derivative, andat least one halogenated compound in a sufficient therapeutic amount tonot substantially stimulate myeloperoxidase activity in a mammal. 14) Amethod according to claim 13, wherein the zwitterionic compound and/orthe amino acid is taurine or a taurine analog. 15) A method according toclaim 13, wherein: the halogenated compound is a hypochlorite ofalkaline metal, and the N-halogenated derivative is N-chlorinated. 16) Amethod according to claim 15, wherein the hypochlorite is sodiumhypochorite. 17) A method according to claim 15, wherein theN-halogenated derivative is N-chlorinated. 18) A method according toclaim 16, wherein the concentration of the sodium hypochlorite isbetween about 6 moles/liter and about 1000.01 femtomoles/liter. 19) Amethod according to claim 14, wherein the concentration of the taurineis between about 5 moles/liter and about 0.01 femtomoles/liter. 20) Amethod for treatment and/or preventing viral infections, and/orbacterial infections, and/or parasitical infections, and/or fungalinfections, and/or diseases generated from non conventionaltransmissible agents, in humans or animals comprising administring to ahuman or animal a pharmaceutically effective amount of a pharmaceuticalcomposition comprising: at least one halogenated compound, and at leastone N-halogenated derivative of at least one compound selected fromzwitterionic compounds and/or the amino acids or their derivatives,without substantial stimulation of myeloperoxidase activity in the humanor animal. 21) A method of treating chronic inflammation, and/orprogressive inflammation, and/or acute inflammation in humans or animalscomprising administering to a human or animal a pharmaceuticallyeffective amount of a pharmaceutical composition comprising: at leastone halogenated compound, and at least one N-halogenated derivative ofat least one compound selected from zwitterionic compounds and/or aminoacids or their derivatives, without substantial stimulation ofmyeloperoxidase activity in the human or animal. 22) A method ofmodulating immunity, in humans or animals comprising administering to ahuman or animal a pharmaceutically effective amount of a pharmaceuticalcomposition comprising: at least one halogenated compound, and at leastone N-halogenated derivative of at least one compound selected fromzwitterionic compounds and/or amino acids or their derivatives, withoutsubstantial stimulatation of myeloperoxidase activity in the human oranimal. 23) A method of stimulating tissue healing in humans or animalscomprising administering to a human or animal a pharmaceuticallyeffective amount of a pharmaceutical composition comprising: at leastone halogenated compound, and at least one N-halogenated derivative ofat least one compound selected from zwitterionic compounds and/or aminoacids or their derivatives, without substantial stimulation ofmyeloperoxidase activity in the human or animal. 24) A method ofpre-surgically, and/or per-surgically, and/or post-surgically irrigatingin humans or animals comprising contacting the surgical site with acomposition comprising: at least one halogenated compound, and at leastone N-halogenated derivative of at least one compound selected fromzwitterionic compounds and/or amino acids or their derivatives, withoutsubstantial stimulation of myeloperoxidase activity in the human oranimal. 25) A method according to claim 20, wherein the compositiontreats lesions and infections linked to periodontitis. 26) A methodaccording to claim 20, wherein the composition treats lesions andinfections linked to herpesviridiae.